Treatment of gliomas using organonitro compound combination therapy

ABSTRACT

The invention provides therapeutic methods and kits for treating a glioma using a particular dosing regimen of the organonitro compound ABDNAZ, radiation therapy, and one of temozolomide, irinotecan, or bevacizumab.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation patent application under 35 U.S.C. §120 of U.S. patent application Ser. No. 16/712,148, filed on Dec. 12,2019, which is a continuation patent application of U.S. patentapplication Ser. No. 15/989,862, filed on May 25, 2018, which is acontinuation patent application of U.S. patent application Ser. No.15/337,378, filed on Oct. 28, 2016, which claims the benefit of andpriority to U.S. Provisional Patent Application No. 62/247,846, filed onOct. 29, 2015, the contents of each of which are hereby incorporated byreference herein, in their entirety.

FIELD OF THE INVENTION

The invention provides therapeutic methods and kits for treating aglioma using a particular dosing regimen of the organonitro compoundABDNAZ, radiation therapy, and one of temozolomide, irinotecan, orbevacizumab.

BACKGROUND

Gliomas are tumors that arise from glial tissue of the brain. Gliomasare often found in the cerebral hemispheres of the brain, but can befound anywhere in the brain. Because gliomas can grow rapidly, commonsymptoms experienced by patients suffering from a glioma are oftenassociated with an increased pressure in the brain. The symptoms caninclude headache, nausea, vomiting, and drowsiness. In addition,patients suffering from a glioma may develop other symptoms such asweakness on one side of the body, memory and/or speech difficulties, andvisual changes, often as function of the location of the glioma tumor.

Gliomas can be characterized according to grade, where low-grade gliomas[WHO grade II] are typically well-differentiated and tend to exhibitbenign tendencies. Low-grade gliomas stand in contrast to high-gradegliomas [WHO grade III or IV] that are typically undifferentiated andfrequently malignant. One example of a high-grade glioma is glioblastomamultiforme. Glioblastoma multiforme has been characterized as a WorldHealth Organization grade IV astrocytoma, with an incidence in NorthAmerica of 5.0 per 100,000 in the population, representing 15 to 20% ofall primary intracranial neoplasms.

Currently available therapeutic approaches for treating patientssuffering from a glioma include surgery, radiation therapy, andtreatment with certain anti-cancer agents. A typical first step intreating a glioblastoma is to surgically remove as much tumor aspossible. This can help alleviate pressure on the brain. However,glioblastomas often have finger-like tentacles, and as a result it canbe difficult to completely remove all the glioblastoma. This isparticularly true when a glioblastoma appears near parts of the brainthat control important functions, such as language and coordination.Radiation and chemotherapy may be used to help slow the growth ofglioblastoma tumors that cannot be removed with surgery. However,survival rates are low using current standard treatments forglioblastoma. For example, using standard treatments currently approvedby government regulatory agencies, the median survival time for adultswith an anaplastic astrocytoma has been reported to be about two tothree years. For children suffering from a high-grade glioma tumor(grade III or IV), the five-year survival rate has been reported to beabout twenty-five percent.

As a result, there is a need for additional therapies to treat patientssuffering from a glioma. The present invention addresses this need andprovides other related advantages.

SUMMARY

The invention provides therapeutic methods and kits for treating aglioma using a particular dosing regimen of the organonitro compoundABDNAZ, radiation therapy, and one of temozolomide, irinotecan, orbevacizumab. The compound ABDNAZ has the following chemical structure:

The therapeutic methods and kits provide a solution to the long unmetneed for a more effective treatment for patients suffering from aglioma. The therapeutic method generally entails (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising temozolomide, irinotecan, or bevacizumab, andwithin a certain amount of time (e.g., about 7 days) subjecting theglioma to radiation therapy, and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the temozolomide, irinotecan, or bevacizumab, in order to treat theglioma. In preferred embodiments, the radiation therapy may beadministered to the patient for a select number of days (e.g., at least2, 3, 4, or 5 days) over a two-week period, or longer duration of timedepending on patient response to the therapy. In preferred embodiments,the formulation comprising ABDNAZ may be administered to the patient fora select number of days per week (e.g., at least 2, 3, or 4 days perweek) over a two-week period, or longer duration of time depending onpatient response to the therapy. The therapy may be used to treatvarious types of gliomas, such as, for example, a primary glioblastomaor a secondary glioblastoma. The invention having been generallydescribed is explained in more detail in the aspects and embodimentsbelow and in the detailed description.

Accordingly, one aspect of the invention provides a method of treating aglioma in a patient. The method comprises the steps of: (a)administering to the patient in need thereof a therapeutically effectiveamount of a formulation comprising temozolomide, and within about 2 daysthereafter subjecting the glioma to radiation therapy; and (b)administering to the patient at least one dose of a therapeuticallyeffective amount of a formulation comprising ABDNAZ within about 7 daysof administration of a first dose of the temozolomide; to treat theglioma.

Another aspect of the invention provides a method of treating a gliomain a patient. The method comprises the steps of: (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising irinotecan, and within about 7 days subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the irinotecan; to treat the glioma.

Another aspect of the invention provides a method of treating a gliomain a patient. The method comprises the steps of: (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising bevacizumab, and within about 7 days subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the bevacizumab; to treat the glioma.

Another aspect of the invention provides a kit for treating a glioma. Incertain embodiments, the kit comprises: (i) a formulation comprisingABDNAZ, and (ii) instructions for treating a glioma according toprocedures described herein, such as (a) administering to the patient inneed thereof a therapeutically effective amount of a formulationcomprising temozolomide, and within about 2 days thereafter subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the temozolomide; to treat the glioma. In an alternative embodiment,the kit comprises: (i) a formulation comprising ABDNAZ, and (ii)instructions for treating a glioma that comprise (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising irinotecan, and within about 7 days subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the irinotecan; to treat the glioma.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph showing response observed using a GBM14 TMZ-Spatient-derived cell line, as described in Example 3.

FIG. 2 is a graph showing response observed using a GBM14 TMZ-Rpatient-derived cell line, as described in Example 3.

FIG. 3 is a graph showing response observed using cells fromtemozolomide-resistant murine EGFRvIII/Arfko neural stem cell-derivedglioma (NSCG) cell line, as described in Example 4.

FIG. 4 is a graph showing percent survival of subjects over time, in theexperiment in which treatment (i.e., administration of ABDNAZ (RRx-001),temozolomide (TMZ), or both ABDNAZ and temozolomide) was started fivedays after intracranial injection of NSCG cells, as described in Example5.

FIG. 5 is a graph showing percent survival of subjects over time, in theexperiment in which treatment (i.e., administration of ABDNAZ (RRx-001),temozolomide (TMZ), or both ABDNAZ and temozolomide) was started tendays after intracranial injection of NSCG cells, as described in Example5.

FIG. 6 is a graph showing the amount of CC3+ cell apoptosis detected inthe assay, as described in Example 6.

FIG. 7 is a graph showing the amount of TMZ (mg) detected per gram oftissue analyzed, as described in Example 7.

FIG. 8 is a graph showing the amount of irinotecan (μg) detected permilligram of tissue analyzed, as described in Example 8.

DETAILED DESCRIPTION

The invention provides therapeutic methods and kits for treating aglioma using a particular dosing regimen of the organonitro compoundABDNAZ, radiation therapy, and one of temozolomide, irinotecan, orbevacizumab. The compound ABDNAZ has the following chemical structure:

The therapeutic methods and kits provide a solution to the long unmetneed for a more effective treatment for patients suffering from aglioma. The therapeutic method generally entails (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising temozolomide, irinotecan, or bevacizumab, andwithin a certain amount of time (e.g., about 7 days) subjecting theglioma to radiation therapy, and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the temozolomide, irinotecan, or bevacizumab, in order to treat theglioma. In preferred embodiments, the radiation therapy may beadministered to the patient for a select number of days (e.g., at least2, 3, 4, or 5 days) over a two-week period, or longer duration of timedepending on patient response to the therapy. In preferred embodiments,the formulation comprising ABDNAZ may be administered to the patient fora select number of days per week (e.g., at least 2, 3, or 4 days perweek) over a two-week period, or longer duration of time depending onpatient response to the therapy. The therapy may be used to treatvarious types of gliomas, such as, for example, a primary glioblastomaor a secondary glioblastoma.

Various aspects of the invention are set forth below in sections;however, aspects of the invention described in one particular sectionare not to be limited to any particular section.

I. Therapeutic Methods for Treating a Glioma

The invention provides therapeutic methods for treating a glioma using aparticular dosing regimen of the organonitro compound ABDNAZ, radiationtherapy, and one of temozolomide, irinotecan, or bevacizumab. Thetherapeutic methods provide a solution to the long unmet need for a moreeffective treatment for patients suffering from a glioma. Thetherapeutic method generally entails (a) administering to the patient inneed thereof a therapeutically effective amount of a formulationcomprising temozolomide, irinotecan, or bevacizumab, and within acertain amount of time (e.g., about 7 days) subjecting the glioma toradiation therapy, and (b) administering to the patient at least onedose of a therapeutically effective amount of a formulation comprisingABDNAZ within about 7 days of administration of a first dose of thetemozolomide, irinotecan, or bevacizumab, in order to treat the glioma.

The invention also provides methods for increasing the amount oftemozolomide, irinotecan, or bevacizumab in a glioma. The methodgenerally involves administering to a patient having a glioma (i) afirst formulation comprising temozolomide, irinotecan, or bevacizumaband (ii) an effective amount of a formulation comprising ABDNAZ so thatthe ABDNAZ exerts physiological activity during a time period in whichthe temozolomide, irinotecan, or bevacizumab is present in the patient,in order to increase the amount of the temozolomide, irinotecan, orbevacizumab in the glioma.

Various features of the methods are described in sections below. Thesections are arranged for convenience and information in one section isnot limited to that section, but may be applied to other sections.

First Method—Using Temozolomide

One aspect of the invention provides a method of treating a glioma in apatient. The method comprises the steps of: (a) administering to thepatient in need thereof a therapeutically effective amount of aformulation comprising temozolomide, and within about 2 days thereaftersubjecting the glioma to radiation therapy; and (b) administering to thepatient at least one dose of a therapeutically effective amount of aformulation comprising ABDNAZ within about 7 days of administration of afirst dose of the temozolomide; to treat the glioma.

Temozolomide is a small molecule having the chemical name3,4-dihydro-3-methyl-4-oxoimidazo[5,1-d]-as-tetrazine-8-carboxamide,which is marketed commercially under the tradename TEMODAR®.

Exemplary Features of the Method

The above method may be further characterized by additional features,such as the dosing schedule and amount of temozolomide, dosing scheduleof ABDNAZ, dosing amount for ABDNAZ, features of the radiation therapy,and the rest period/maintenance period.

Dosing Schedule and Amount of Temozolomide

The method can be further characterized according to the dosing scheduleand amount of the temozolomide. Accordingly, in certain embodiments, theformulation comprising temozolomide is administered daily for at least 2weeks. In certain embodiments, the formulation comprising temozolomideis administered daily for at least 4 weeks. In certain embodiments, theformulation comprising temozolomide is administered daily for at least 6weeks. In certain embodiments, the formulation comprising temozolomideis administered at the patient's bedtime.

In certain embodiments, the patient receives temozolomide by oraladministration at a daily dose of at least 50 mg/m². In certainembodiments, the patient receives temozolomide by oral administration ata daily dose of about 50 mg/m² to about 100 mg/m². In certainembodiments, the patient receives temozolomide by oral administration ata daily dose of about 75 mg/m².

Dosing Schedule of ABDNAZ

A formulation comprising ABDNAZ may be administered multiple times, suchas multiple times over a defined period of time. Further, coordinationof the dosing schedule of the temozolomide and radiation therapy withthat of the formulation comprising ABDNAZ is contemplated to providetherapeutic benefits, such as superior efficacy.

In certain embodiments, for a duration of at least 2 weeks followingadministration of the first dose of temozolomide, the patient receivesat least one dose each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, for a duration ofat least 4 weeks following administration of the first dose oftemozolomide, the patient receives at least one dose each week of atherapeutically effective amount of a formulation comprising ABDNAZ. Incertain embodiments, for a duration of at least 6 weeks followingadministration of the first dose of temozolomide, the patient receivesat least one dose each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, two doses of atherapeutically effective amount of a formulation comprising ABDNAZ areadministered within about 7 days of administration of the first dose oftemozolomide. In certain embodiments, for a duration of at least 2 weeksfollowing administration of the first dose of temozolomide, the patientreceives two doses each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, for a duration ofat least 4 weeks following administration of the first dose oftemozolomide, the patient receives two doses each week of atherapeutically effective amount of a formulation comprising ABDNAZ. Incertain embodiments, for a duration of at least 6 weeks followingadministration of the first dose of temozolomide, the patient receivestwo doses each week of a therapeutically effective amount of aformulation comprising ABDNAZ.

In certain embodiments, for a duration of at least 2 weeks followingadministration of the first dose of temozolomide, the patient receivesone dose each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, for a duration ofat least 4 weeks following administration of the first dose oftemozolomide, the patient receives one dose each week of atherapeutically effective amount of a formulation comprising ABDNAZ. Incertain embodiments, for a duration of at least 6 weeks followingadministration of the first dose of temozolomide, the patient receivesone dose each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, for a duration of6 weeks following administration of the first dose of temozolomide, thepatient receives one dose each week of a therapeutically effectiveamount of a formulation comprising ABDNAZ.

In certain embodiments, any dose of ABDNAZ administered to the patientwithin 12 hours of administering temozolomide is administered at least 3hours before administering the temozolomide.

In certain embodiments, the patient receives ABDNAZ within about 3 hoursprior to subjecting the glioma to radiation therapy. In certainembodiments, the patient receives ABDNAZ within about 6 hours prior tosubjecting the glioma to radiation therapy. In certain embodiments, thepatient receives ABDNAZ within about 24 hours prior to subjecting theglioma to radiation therapy. In certain embodiments, the patientreceives ABDNAZ within about 48 hours prior to subjecting the glioma toradiation therapy.

In certain embodiments, the formulation comprising ABDNAZ isadministered so that the ABDNAZ exerts physiological activity during anoverlapping time period with one or both of the radiation therapy andtemozolomide.

Dosing Amount of ABDNAZ

The method may be further a characterized according to the dose ofABDNAZ administered to the patient. The dose of ABDNAZ described hereinfor use in combination with temozolomide and the radiation therapy hasbeen selected in view of the dosing schedule and amount of temozolomideand the radiation therapy. Dosing amounts of ABDNAZ are providedaccording to the number of milligrams of ABDNAZ to be administered tothe patient based on the surface area of the patient as measured in m².

In certain embodiments, each dose of the formulation comprising ABDNAZis administered to the patient by intravenous infusion. In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2 mg/m² to about 20 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2.5 mg/m² to about 5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 5 mg/m² to about 10 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 10 mg/m² to about 16.5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 2.5 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 5 mg/m². Incertain embodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 10 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 16.5 mg/m².

In certain embodiments, each dose of the formulation comprising ABDNAZis administered to the patient by intravenous infusion providing ABDNAZin an amount of from about 0.1 mg to about 20 mg. In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of from about 0.1 mg to about 10 mg. In certain embodiments,each dose of the formulation comprising ABDNAZ is administered to thepatient by intravenous infusion providing ABDNAZ in an amount of fromabout 0.5 mg to about 4.0 mg. In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 0.5 mg, 1.0mg, 1.5 mg, 2.0 mg, 2.5 mg, 3.0 mg, 3.5 mg, or 4.0 mg.

Radiation Therapy Features

The method can be further characterized according to the dosing scheduleand amount of radiation administered for the radiation therapy.

In certain embodiments, the glioma is subjected to radiation therapyonce per day for at least 3 days within a 7 day period followingadministration of the first dose of temozolomide. In certainembodiments, the glioma is subjected to radiation therapy once per dayfor at least 5 days within a 7 day period following administration ofthe first dose of temozolomide. In certain embodiments, for a durationof at least 2 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 3 days of each week. In certain embodiments, for a durationof at least 4 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 3 days of each week. In certain embodiments, for a durationof at least 6 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 3 days of each week. In certain embodiments, for a durationof at least 2 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 5 days of each week. In certain embodiments, for a durationof at least 4 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 5 days of each week. In certain embodiments, for a durationof at least 6 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor at least 5 days of each week. In certain embodiments, for a durationof at least 6 weeks following administration of the first dose oftemozolomide, the glioma is subjected to radiation therapy once per dayfor 5 days each week. In certain embodiments, for a duration of 6 weeksfollowing administration of the first dose of temozolomide, the gliomais subjected to radiation therapy once per day for 5 days each week.

In certain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is from about 1 Gy to about 3 Gy. Incertain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is about 2 Gy. In certainembodiments, the glioma is exposed to from about 50 Gy to about 70 Gy ofradiation by the radiation therapy over a period of 6 weeks followingadministration of the first dose of temozolomide. In certainembodiments, the glioma is exposed to about 60 Gy of radiation by theradiation therapy over a period of 6 weeks following administration ofthe first dose of temozolomide.

In certain embodiments, the radiation therapy is (i) conventionalfractionated external beam radiation or (ii) intensity-modulatedradiation therapy. In certain embodiments, the radiation therapy isconventional fractionated external beam radiation.

Various types of radiation therapy are used by those skilled in the artand have been described in the literature. Exemplary types of radiationtherapy include, for example, radiation therapy comprising gamma rays,X-rays, electron beams, neutron beams, particulate radiation, protonbeams, or the like. The source of the radiation is desirably external tothe patient, which involves directing a beam of high-energy radiation tothe glioma using a machine external to the patient. Desirably the targetsite (i.e., site of the glioma) is exposed to the radiation therapy fora short duration of time, such as less than about 3 hours, 2 hours, 1hour, 30 minutes, 15 minutes, 10 minutes, 5 minutes, or 1 minute foreach dose of radiation therapy.

Rest Period/Maintenance Period

The method may be further characterized according to the scope of therest period/maintenance period. For example, in certain embodiments,after receiving temozolomide for a duration of 6 weeks, the patient doesnot receive any of temozolomide, radiation therapy, or ABDNAZ for aduration of at least about 2 weeks.

In certain embodiments, the patient after having not received any oftemozolomide, radiation therapy, or ABDNAZ for a duration of at leastabout 2 weeks, then receives (i) a therapeutically effective amount of aformulation comprising temozolomide on at least three days during thefirst week, and (ii) a therapeutically effective amount of a formulationcomprising ABDNAZ at least once per week for a duration of at least twoweeks. In certain embodiments, the patient after having not received anyof temozolomide, radiation therapy, or ABDNAZ for a duration of at leastabout 2 weeks, then receives (i) a therapeutically effective amount of aformulation comprising temozolomide on at least five days during thefirst week, and (ii) a therapeutically effective amount of a formulationcomprising ABDNAZ at least once per week for a duration of at least 1month. In certain embodiments, the patient after having not received anyof temozolomide, radiation therapy, or ABDNAZ for a duration of at leastabout 2 weeks, then receives (i) a therapeutically effective amount of aformulation comprising temozolomide on at least five consecutive daysduring the first week of each 28 day period, and (ii) a therapeuticallyeffective amount of a formulation comprising ABDNAZ at least once perweek for a duration of at least 4 months. In certain embodiments, afterreceiving temozolomide for a duration of 6 weeks, the patient does notreceive any of temozolomide, radiation therapy, or ABDNAZ for a durationof at least about 4 weeks. In certain embodiments, the patient afterhaving not received any of temozolomide, radiation therapy, or ABDNAZfor a duration of at least about 4 weeks, then receives (i) atherapeutically effective amount of a formulation comprisingtemozolomide on at least three days during the first week, and (ii) atherapeutically effective amount of a formulation comprising ABDNAZ atleast once per week for a duration of at least two weeks. In certainembodiments, the patient after having not received any of temozolomide,radiation therapy, or ABDNAZ for a duration of at least about 4 weeks,then receives (i) a therapeutically effective amount of a formulationcomprising temozolomide on at least five days during the first week, and(ii) a therapeutically effective amount of a formulation comprisingABDNAZ at least once per week for a duration of at least 1 month. Incertain embodiments, the patient after having not received any oftemozolomide, radiation therapy, or ABDNAZ for a duration of at leastabout 4 weeks, then receives (i) a therapeutically effective amount of aformulation comprising temozolomide on at least five consecutive daysduring the first week of each 28 day period, and (ii) a therapeuticallyeffective amount of a formulation comprising ABDNAZ at least once perweek for a duration of at least 4 months.

In certain embodiments, after receiving temozolomide for a duration of 6weeks, the patient does not receive any of temozolomide, radiationtherapy, or ABDNAZ for a duration of about 3 weeks to about 6 weeks.

In certain embodiments, after the duration over which the patient hasnot received any of temozolomide, radiation therapy, or ABDNAZ, theformulation comprising temozolomide is administered orally to thepatient to provide temozolomide in an amount of at least 150 mg/m². Incertain embodiments, after the duration over which the patient has notreceived any of temozolomide, radiation therapy, or ABDNAZ, theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 5 mg/m². Incertain embodiments, after the duration over which the patient has notreceived any of temozolomide, radiation therapy, or ABDNAZ, theformulation comprising temozolomide is administered orally to thepatient to provide temozolomide in an amount of from about 75 mg/m² toabout 400 mg/m². In certain embodiments, after the duration over whichthe patient has not received any of temozolomide, radiation therapy, orABDNAZ, the formulation comprising temozolomide is administered orallyto the patient to provide temozolomide in an amount of from about 150mg/m² to about 350 mg/m². In certain embodiments, after the durationover which the patient has not received any of temozolomide, radiationtherapy, or ABDNAZ, the formulation comprising temozolomide isadministered orally to the patient to provide temozolomide in an amountof about 150 mg/m², 200 mg/m², 250 mg/m², 300 mg/m², or 350 mg/m².

In certain embodiments, after the duration over which the patient hasnot received any of temozolomide, radiation therapy, or ABDNAZ, theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of from about 0.1 mgto about 10 mg. In certain embodiments, after the duration over whichthe patient has not received any of temozolomide, radiation therapy, orABDNAZ, the formulation comprising ABDNAZ is administered to the patientby intravenous infusion providing ABDNAZ in an amount of from about 0.5mg to about 4.0 mg. In certain embodiments, after the duration overwhich the patient has not received any of temozolomide, radiationtherapy, or ABDNAZ, the formulation comprising ABDNAZ is administered tothe patient by intravenous infusion providing ABDNAZ in an amount ofabout 0.5 mg, 1.0 mg, 1.5, mg, 2.0 mg, 2.5 mg, 3.0 mg, or 4.0 mg.

Combinations of Embodiments and Illustration of Exemplary More SpecificMethods

All combinations of aspects and embodiments described above arecontemplated. For example, a therapeutic method is contemplated fortreating a glioblastoma by administering to a patient in need thereof(a) for a duration of at least 3 weeks, orally administer one dose of aformulation comprising temozolimide to the patient daily, intravenouslyadminister a formulation comprising ABDNAZ at least once per week, andsubject the glioma to radiation therapy at least 3 days each week, andthereafter (b) do not administer any temozolimide, ABDNAZ, or radiationtherapy to the patient for at least two weeks, and thereafter (c) orallyadminister one dose of a formulation comprising temozolimide to thepatient at least 3 days each month, and intravenously administer aformulation comprising ABDNAZ at least once per week.

Exemplary more specific illustrations of the contemplated dosing methodsfor treating a glioma (e.g., a glioblastoma) are provided below inTables 1, 2, 3, and 4.

TABLE 1 Step No. Dosing Schedule 1 For a duration of at least 3 weeks,orally administer one dose of a formulation comprising temozolimide tothe patient daily, intravenously administer a formulation comprisingABDNAZ at least once per week, and subject the glioma to radiationtherapy at least 3 days each week. 2 After completing step 1, do notadminister any temozolimide, ABDNAZ, or radiation therapy to the patientfor at least two weeks. 3 After completing step 2, orally administer onedose of a formulation comprising temozolimide to the patient at least 3days each month, and intravenously administer a formulation comprisingABDNAZ at least once per week.

TABLE 2 Step No. Dosing Schedule 1 For a duration of at least 3 weeks,orally administer one dose of a formulation comprising temozolimide tothe patient daily at a daily dosage of at least 75 mg/m², intravenouslyadminister a formulation comprising ABDNAZ at least once per week, andsubject the glioma to radiation therapy at least 3 days each week. 2After completing step 1, do not administer any temozolimide, ABDNAZ, orradiation therapy to the patient for at least two weeks. 3 Aftercompleting step 2, orally administer one dose of a formulationcomprising temozolimide to the patient at least 3 days each month, andintravenously administer a formulation comprising ABDNAZ at least onceper week.

TABLE 3 Step No. Dosing Schedule 1 For a duration of at least 6 weeks,orally administer one dose of a formulation comprising temozolimide tothe patient daily at a daily dosage of at least 75 mg/m², intravenouslyadminister one dose of a formulation comprising ABDNAZ at least twiceonce per week wherein each dose of the formulation comprising ABDNAZprovides from about 2.5 mg/m² to about 16.5 mg/m² of ABDNAZ, and subjectthe glioma to a dose of radiation therapy at least 5 days each weekwherein each dose of radiation therapy provides from about 1 Gy to about3 Gy of radiation. 2 After completing step 1, do not administer anytemozolimide, ABDNAZ, or radiation therapy to the patient for at leastfour weeks. 3 After completing step 2, for a duration of at least twomonths orally administering one dose of a formulation comprisingtemozolimide to the patient at least 5 days each month wherein each doseof the formulation comprising temozolimide provides from about 150 mg/m²to about 200 mg/m² of temozolimide, and intravenously administer a doseof a formulation comprising ABDNAZ at least once per week where eachdose of the formulation comprising ABDNAZ provides at least 5 mg/m² ofABDNAZ.

TABLE 4 Step No. Dosing Schedule 1 For a duration of at about 6 weeks,orally administer one dose of a formulation comprising temozolimide tothe patient daily at a daily dosage of at least 75 mg/m², intravenouslyadminister one dose of a formulation comprising ABDNAZ twice once perweek wherein each dose of the formulation comprising ABDNAZ providesfrom about 2.5 mg/m² to about 16.5 mg/m2 of ABDNAZ, and subject theglioma to a dose of radiation therapy 5 days each week wherein each doseof radiation therapy provides about 2 Gy of radiation. 2 Aftercompleting step 1, do not administer any temozolimide, ABDNAZ, orradiation therapy to the patient for about four weeks. 3 Aftercompleting step 2, for a duration of at least two months orallyadministering one dose of a formulation comprising temozolimide to thepatient about 5 days each month wherein each dose of the formulationcomprising temozolimide provides from about 150 mg/m² to about 200 mg/m²of temozolimide, and intravenously administer a dose of a formulationcomprising ABDNAZ once per week where each dose of the formulationcomprising ABDNAZ provides about 5 mg/m² of ABDNAZ.

A further exemplary more specific dosing method for treating a glioma(e.g., a glioblastoma) is as follows:

-   -   Step 1: for a duration of at about 6 weeks, orally administer        one dose of a formulation comprising temozolimide to the patient        daily at a daily dosage of about 75 mg/m², intravenously        administer one dose of a formulation comprising ABDNAZ once per        week wherein each dose of the formulation comprising ABDNAZ        provides from about 0.5 mg to about 4.0 mg of ABDNAZ, and        subject the glioma to a dose of radiation therapy 5 days each        week wherein each dose of radiation therapy provides about 2 Gy        of radiation. The ABDNAZ is administered within 3 hours prior to        administering radiation therapy, and the temozolomide is        administered no sooner than 3 hours after administering the        ABDNAZ.    -   Step 2: after completing step 1, do not administer any        temozolimide, ABDNAZ, or radiation therapy to the patient for        about three to six weeks.    -   Step 3: after completing step 2, for a duration of at least five        months (preferably for six months) orally administer one dose of        a formulation comprising temozolimide to the patient about 5        days each month (e.g., days 1-5 of each month) wherein each dose        of the formulation comprising temozolimide provides from about        150 mg/m² to about 350 mg/m² of temozolimide, and intravenously        administer one dose of a formulation comprising ABDNAZ once per        week where each dose of the formulation comprising ABDNAZ        provides about from about 0.5 mg to about 4.0 mg of ABDNAZ.

Second Method—Using Irinotecan

Another aspect of the invention provides a method of treating a gliomain a patient. The method comprises the steps of: (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising irinotecan, and within about 7 days subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the irinotecan; to treat the glioma.

Irinotecan is commercially available and marketed in the form of amonohydrochloride trihydrate under the tradename CAMPTOSAR®.

Exemplary Features of the Method

The above method may be further characterized by additional features,such as the dosing schedule and amount of irinotecan, dosing schedule ofABDNAZ, dosing amount for ABDNAZ, features of the radiation therapy, andform of irinotecan.

Dosing Schedule and Amount of Irinotecan

The method can be further characterized according to the dosing scheduleand amount of the irinotecan. Accordingly, in certain embodiments, theformulation comprising irinotecan is administered at least once per weekfor a duration of at least 2 weeks. In certain embodiments, theformulation comprising irinotecan is administered at least once per weekfor a duration of at least 4 weeks.

In certain embodiments, the patient receives the formulation comprisingirinotecan by (i) intravenous infusion once weekly at a single dose ofat least about 100 mg/m² of irinotecan, (ii) intravenous infusion onceevery two weeks at a single dose of at least about 200 mg/m² ofirinotecan, or (iii) intravenous infusion once every three weeks at asingle dose of at least about 300 mg/m² of irinotecan. In certainembodiments, the patient receives the formulation comprising irinotecanby intravenous infusion once weekly at a single dose of about 125 mg/m²of irinotecan.

Dosing Schedule for ABDNAZ

A formulation comprising ABDNAZ may be administered multiple times, suchas multiple times over a defined period of time. Further, coordinationof the dosing schedule of the irinotecan and radiation therapy with thatof the formulation comprising ABDNAZ is contemplated to providetherapeutic benefits, such as superior efficacy.

In certain embodiments, two doses of a therapeutically effective amountof a formulation comprising ABDNAZ are administered within about 7 daysof administration of the first dose of irinotecan. In certainembodiments, for a duration of at least 2 weeks following administrationof the first dose of irinotecan, the patient receives two doses eachweek of a therapeutically effective amount of a formulation comprisingABDNAZ. In certain embodiments, for a duration of at least 4 weeksfollowing administration of the first dose of irinotecan, the patientreceives two doses each week of a therapeutically effective amount of aformulation comprising ABDNAZ.

In certain embodiments, for a duration of at least 2 weeks followingadministration of the first dose of irinotecan, the patient receives onedose each week of a therapeutically effective amount of a formulationcomprising ABDNAZ. In certain embodiments, for a duration of at least 4weeks following administration of the first dose of irinotecan, thepatient receives one dose each week of a therapeutically effectiveamount of a formulation comprising ABDNAZ. In certain embodiments, for aduration of at least 6 weeks following administration of the first doseof irinotecan, the patient receives one dose each week of atherapeutically effective amount of a formulation comprising ABDNAZ. Incertain embodiments, for a duration of 6 weeks following administrationof the first dose of irinotecan, the patient receives one dose each weekof a therapeutically effective amount of a formulation comprisingABDNAZ.

In certain embodiments, the patient receives ABDNAZ within about 6 hoursprior to subjecting the glioma to radiation therapy. In certainembodiments, the patient receives ABDNAZ within about 24 hours prior tosubjecting the glioma to radiation therapy. In certain embodiments, thepatient receives ABDNAZ within about 48 hours prior to subjecting theglioma to radiation therapy.

In certain embodiments, the formulation comprising ABDNAZ isadministered so that the ABDNAZ exerts physiological activity during anoverlapping time period with one or both of the radiation therapy andirinotecan.

Dose of ABDNAZ Administered

The dose ABDNAZ described herein for use in combination with irinotecanand radiation therapy has been selected in view of the dosing amount anddosing schedule of the irinotecan and radiation therapy. Dosing amountsof ABDNAZ are provided according to the number of milligrams of ABDNAZto be administered to the patient based on the surface area of thepatient as measured in m².

In certain embodiments, each dose of the formulation comprising ABDNAZis administered to the patient by intravenous infusion. In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2 mg/m² to about 20 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2.5 mg/m² to about 5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 5 mg/m² to about 10 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 10 mg/m² to about 16.5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 2.5 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 5 mg/m². Incertain embodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 10 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 16.5 mg/m².

In certain embodiments, each dose of the formulation comprising ABDNAZis administered to the patient by intravenous infusion providing ABDNAZin an amount of from about 0.1 mg to about 20 mg. In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of from about 0.1 mg to about 10 mg. In certain embodiments,each dose of the formulation comprising ABDNAZ is administered to thepatient by intravenous infusion providing ABDNAZ in an amount of fromabout 0.5 mg to about 4.0 mg. In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 0.5 mg, 1.0mg, 1.5 mg, 2.0 mg, 2.5 mg, 3.0 mg, 3.5 mg, or 4.0 mg.

Radiation Therapy Features

In certain embodiments, the glioma is subjected to radiation therapyonce per day for at least 3 days within a 7 day period followingadministration of the first dose of irinotecan. In certain embodiments,the glioma is subjected to radiation therapy once per day for at least 5days within a 7 day period following administration of the first dose ofirinotecan. In certain embodiments, for a duration of at least 2 weeksfollowing administration of the first dose of irinotecan, the glioma issubjected to radiation therapy once per day for at least 5 days of eachweek. In certain embodiments, for a duration of at least 4 weeksfollowing administration of the first dose of irinotecan, the glioma issubjected to radiation therapy once per day for at least 5 days of eachweek.

In certain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is from about 1 Gy to about 3 Gy. Incertain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is about 2 Gy.

In certain embodiments, the glioma is exposed to from about 50 Gy toabout 70 Gy of radiation by the radiation therapy over a period of 6weeks following administration of the first dose of irinotecan. Incertain embodiments, the glioma is exposed to about 60 Gy of radiationby the radiation therapy over a period of 6 weeks followingadministration of the first dose of irinotecan.

In certain embodiments, the radiation therapy is (i) conventionalfractionated external beam radiation or (ii) intensity-modulatedradiation therapy. In certain embodiments, the radiation therapy is (i)conventional fractionated external beam radiation.

Various types of radiation therapy are used by those skilled in the artand have been described in the literature. Exemplary types of radiationtherapy include, for example, radiation therapy comprising gamma rays,X-rays, electron beams, neutron beams, particulate radiation, protonbeams, or the like. The source of the radiation is desirably external tothe patient, which involves directing a beam of high-energy radiation tothe glioma using a machine external to the patient. Desirably the targetsite (i.e., site of the glioma) is exposed to the radiation therapy fora short duration of time, such as less than about 3 hours, 2 hours, 1hour, 30 minutes, 15 minutes, 10 minutes, 5 minutes, or 1 minute foreach dose of radiation therapy.

Form of Irinotecan

The method may be further characterized according to the form ofirinotecan used, such as a free base or pharmaceutically acceptable salt(such an acid addition salt). In certain embodiments, the formulationcomprising irinotecan comprises irinotecan hydrochloride.

Combinations of Embodiments and Illustration of Exemplary More SpecificMethods

All combinations of aspects and embodiments described above arecontemplated. For example, a therapeutic method is contemplated fortreating a glioblastoma by administering to a patient in need thereof(a) for a duration of at least 3 weeks, orally administer one dose of aformulation comprising irinotecan to the patient daily, intravenouslyadminister a formulation comprising ABDNAZ at least once per week, andsubject the glioma to radiation therapy at least 3 days each week, andthereafter (b) do not administer any irinotecan, ABDNAZ, or radiationtherapy to the patient for at least two weeks, and thereafter (c) orallyadminister one dose of a formulation comprising irinotecan to thepatient at least 3 days each month, and intravenously administer aformulation comprising ABDNAZ at least once per week.

Exemplary more specific illustrations of the contemplated dosing methodsfor treating a glioma (e.g., a glioblastoma) are provided below inTables 5, 6, 7, 8, and 9.

TABLE 5 Step No. Dosing Schedule 1 For a duration of at least 2 weeks,intravenously administer one dose of a formulation comprising irinotecanto the patient at least once per week wherein each dose provides atleast 100 mg/m² of irinotecan, intravenously administer a formulationcomprising ABDNAZ at least once per week, and subject the glioma toradiation therapy at least once per week. 2 After completing step 1, donot administer any irinotecan to the patient for at least one week.

TABLE 6 Step No. Dosing Schedule 1 For a duration of at least 4 weeks,intravenously administer one dose of a formulation comprising irinotecanto the patient at least once per week wherein each dose provides atleast 100 mg/m² of irinotecan, intravenously administer a formulationcomprising ABDNAZ at least once per week, and subject the glioma toradiation therapy at least twice per week. 2 After completing step 1, donot administer any irinotecan to the patient for at least two weeks.

TABLE 7 Step No. Dosing Schedule 1 For a duration of at least 2 weeks,intravenously administer one dose of a formulation comprising irinotecanto the patient at least once per week wherein each dose provides atleast 125 mg/m² of irinotecan, intravenously administer one dose of aformulation comprising ABDNAZ at least twice once per week wherein eachdose of the formulation comprising ABDNAZ provides from about 2.5 mg/m²to about 16.5 mg/m² of ABDNAZ, and subject the glioma to a dose ofradiation therapy at least 2 days each week wherein each dose ofradiation therapy provides from about 1 Gy to about 3 Gy of radiation. 2After completing step 1, do not administer any irinotecan for at leastone week.

TABLE 8 Step No. Dosing Schedule 1 For a duration of at least 4 weeks,intravenously administer one dose of a formulation comprising irinotecanto the patient once per week wherein each dose provides at least 125mg/m² of irinotecan, intravenously administer one dose of a formulationcomprising ABDNAZ at least twice once per week wherein each dose of theformulation comprising ABDNAZ provides from about 2.5 mg/m² to about16.5 mg/m² of ABDNAZ, and subject the glioma to a dose of radiationtherapy at least 2 days each week wherein each dose of radiation therapyprovides from about 1 Gy to about 3 Gy of radiation. 2 After completingstep 1, do not administer any irinotecan for at least one week.

TABLE 9 Step No. Dosing Schedule 1 For a duration of at least 3 weeks,intravenously administer one dose of a formulation comprising irinotecanto the patient once every three weeks wherein each dose provides atleast 350 mg/m² of irinotecan, intravenously administer one dose of aformulation comprising ABDNAZ at least twice once per week wherein eachdose of the formulation comprising ABDNAZ provides from about 2.5 mg/m²to about 16.5 mg/m² of ABDNAZ, and subject the glioma to a dose ofradiation therapy at least 2 days each week wherein each dose ofradiation therapy provides from about 1 Gy to about 3 Gy of radiation. 2After completing step 1, do not administer any irinotecan for at leasttwo weeks.

Third Method—Using Bevacizumab

Another aspect of the invention provides a method of treating a gliomain a patient. The method comprises the steps of: (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising bevacizumab, and within about 7 days subjectingthe glioma to radiation therapy; and (b) administering to the patient atleast one dose of a therapeutically effective amount of a formulationcomprising ABDNAZ within about 7 days of administration of a first doseof the bevacizumab; to treat the glioma.

Bevacizumab is a recombinant humanized monoclonal IgG1 antibody thatbinds to and inhibits the biologic activity of human vascularendothelial growth factor (VEGF) in in vitro and in vivo assay systems.Bevacizumab contains human framework regions and thecomplementarity-determining regions of a murine antibody that binds toVEGF. Bevacizumab has an approximate molecular weight of 149 kD and canbe produced in a mammalian cell (Chinese Hamster Ovary) expressionsystem in a nutrient medium containing the antibiotic gentamicin.Bevacizumab is marketed commercially as AVASTIN®. The formulation ofbevacizumab marketed commercially as AVASTIN® is a clear to slightlyopalescent, colorless to pale brown, sterile, pH 6.2 solution forintravenous infusion. The formulation is supplied in 100 mg and 400 mgamounts of bevacizumab in preservative-free, single-use vials. The 100mg product is formulated in 240 mg α,α-trehalose dihydrate, 23.2 mgsodium phosphate (monobasic, monohydrate), 4.8 mg sodium phosphate(dibasic, anhydrous), 1.6 mg polysorbate 20, and Water for Injection,USP. The 400 mg product is formulated in 960 mg α,α-trehalose dihydrate,92.8 mg sodium phosphate (monobasic, monohydrate), 19.2 mg sodiumphosphate (dibasic, a 695 nhydrous), 6.4 mg polysorbate 20, and Waterfor Injection, USP.

Exemplary Features of the Method

The above method may be further characterized by additional features,such as the dosing schedule and amount of bevacizumab, dosing scheduleof ABDNAZ, dosing amount for ABDNAZ, and features of the radiationtherapy.

Dosing Schedule and Amount of Bevacizumab

The method can be further characterized according to the dosing scheduleand amount of the bevacizumab. Accordingly, in certain embodiments, theformulation comprising bevacizumab is administered once every 2 weeksfor a duration of at least 4 weeks. In certain embodiments, theformulation comprising bevacizumab is administered once every 3 weeksfor a duration of at least 6 weeks.

In certain embodiments, the patient receives the formulation comprisingbevacizumab by (i) intravenous infusion once every 2 weeks at a dose ofat least about 5 mg/kg of bevacizumab for a duration of at least 4, 6,8, 10, or 12 weeks, (ii) intravenous infusion once every 2 weeks at adose of at least about 10 mg/kg of bevacizumab for a duration of atleast 4, 6, 8, 10, or 12 weeks, or (iii) intravenous infusion once every3 weeks at a dose of at least about 15 mg/kg of bevacizumab for aduration of at least 6, 12, 15, or 18 weeks. In certain embodiments, thepatient receives the formulation comprising bevacizumab by intravenousinfusion once every 2 weeks at a dose of 5-10 mg/kg of bevacizumab for aduration of at least 4, 6, 8, 10, or 12 weeks.

In certain other embodiments, the patient receives the formulationcomprising bevacizumab by (i) intravenous infusion once every 2 weeks ata dose of about 5 mg/kg of bevacizumab for a duration of at least 4, 6,8, 10, or 12 weeks, (ii) intravenous infusion once every 2 weeks at adose of about 10 mg/kg of bevacizumab for a duration of at least 4, 6,8, 10, or 12 weeks, or (iii) intravenous infusion once every 3 weeks ata dose of about 15 mg/kg of bevacizumab for a duration of at least 6,12, 15, or 18 weeks. In certain embodiments, the patient receives theformulation comprising bevacizumab by intravenous infusion once every 2weeks at a dose of 5-20 mg/kg of bevacizumab for a duration of at least4, 6, 8, 10, or 12 weeks.

Dosing Schedule for ABDNAZ

A formulation comprising ABDNAZ may be administered multiple times, suchas multiple times over a defined period of time. Further, coordinationof the dosing schedule of the bevacizumab and radiation therapy withthat of the formulation comprising ABDNAZ is contemplated to providetherapeutic benefits, such as superior efficacy.

In certain embodiments, two doses of a therapeutically effective amountof a formulation comprising ABDNAZ are administered within about 7 daysof administration of the first dose of bevacizumab. In certainembodiments, for a duration of at least 2 weeks following administrationof the first dose of bevacizumab, the patient receives two doses eachweek of a therapeutically effective amount of a formulation comprisingABDNAZ. In certain embodiments, for a duration of at least 4 weeksfollowing administration of the first dose of bevacizumab, the patientreceives two doses each week of a therapeutically effective amount of aformulation comprising ABDNAZ. In certain embodiments, the patientreceives ABDNAZ within about 6 hours prior to subjecting the glioma toradiation therapy. In certain embodiments, the patient receives ABDNAZwithin about 24 hours prior to subjecting the glioma to radiationtherapy. In certain embodiments, the patient receives ABDNAZ withinabout 48 hours prior to subjecting the glioma to radiation therapy.

In certain embodiments, the formulation comprising ABDNAZ isadministered so that the ABDNAZ exerts physiological activity during anoverlapping time period with one or both of the radiation therapy andbevacizumab.

Dose of ABDNAZ Administered

The dose ABDNAZ described herein for use in combination with bevacizumaband radiation therapy has been selected in view of the dosing amount anddosing schedule of the bevacizumab and radiation therapy. Dosing amountsof ABDNAZ are provided according to the number of milligrams of ABDNAZto be administered to the patient based on the surface area of thepatient as measured in m².

In certain embodiments, each dose of the formulation comprising ABDNAZis administered to the patient by intravenous infusion. In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2 mg/m² to about 20 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 2.5 mg/m² to about 5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 5 mg/m² to about 10 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount ranging from about 10 mg/m² to about 16.5 mg/m². In certainembodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 2.5 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 5 mg/m². Incertain embodiments, each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of about 10 mg/m². In certain embodiments, each dose of theformulation comprising ABDNAZ is administered to the patient byintravenous infusion providing ABDNAZ in an amount of about 16.5 mg/m².

Radiation Therapy Features

In certain embodiments, the glioma is subjected to radiation therapyonce per day for at least 3 days within a 7 day period followingadministration of the first dose of bevacizumab. In certain embodiments,the glioma is subjected to radiation therapy once per day for at least 5days within a 7 day period following administration of the first dose ofbevacizumab. In certain embodiments, for a duration of at least 2 weeksfollowing administration of the first dose of bevacizumab, the glioma issubjected to radiation therapy once per day for at least 5 days of eachweek. In certain embodiments, for a duration of at least 4 weeksfollowing administration of the first dose of bevacizumab, the glioma issubjected to radiation therapy once per day for at least 5 days of eachweek.

In certain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is from about 1 Gy to about 3 Gy. Incertain embodiments, when radiation therapy is administered to theglioma, the amount of radiation provided to the glioma on the day ofadministering the radiation therapy is about 2 Gy.

In certain embodiments, the glioma is exposed to from about 50 Gy toabout 70 Gy of radiation by the radiation therapy over a period of 6weeks following administration of the first dose of bevacizumab. Incertain embodiments, the glioma is exposed to about 60 Gy of radiationby the radiation therapy over a period of 6 weeks followingadministration of the first dose of bevacizumab.

In certain embodiments, the radiation therapy is (i) conventionalfractionated external beam radiation or (ii) intensity-modulatedradiation therapy. In certain embodiments, the radiation therapy is (i)conventional fractionated external beam radiation.

Various types of radiation therapy are used by those skilled in the artand have been described in the literature. Exemplary types of radiationtherapy include, for example, radiation therapy comprising gamma rays,X-rays, electron beams, neutron beams, particulate radiation, protonbeams, or the like. The source of the radiation is desirably external tothe patient, which involves directing a beam of high-energy radiation tothe glioma using a machine external to the patient. Desirably the targetsite (i.e., site of the glioma) is exposed to the radiation therapy fora short duration of time, such as less than about 3 hours, 2 hours, 1hour, 30 minutes, 15 minutes, 10 minutes, 5 minutes, or 1 minute foreach dose of radiation therapy.

Combinations of Embodiments and Illustration of Exemplary More SpecificMethods

All combinations of aspects and embodiments described above arecontemplated. For example, a therapeutic method is contemplated fortreating a glioblastoma by administering to a patient in need thereoffor a duration of at least 3 weeks, orally administer one dose of aformulation comprising bevacizumab to the patient daily, intravenouslyadminister a formulation comprising ABDNAZ at least once per week, andsubject the glioma to radiation therapy at least 3 days each week.Exemplary more specific illustrations of the contemplated dosing methodsfor treating a glioma (e.g., a glioblastoma) using bevacizumab areprovided below in Table 10 below describing multiple more specificdosing schedules.

TABLE 10 Dosing Description of Schedule Dosing Schedule A For a durationof at least 4 weeks, intravenously administer one dose of a formulationcomprising bevacizumab to the patient once every two weeks wherein eachdose provides at least 5 mg/kg of bevacizumab, intravenously administera formulation comprising ABDNAZ at least once per week, and subject theglioma to radiation therapy at least once per week. B For a duration ofat least 4 weeks, intravenously administer one dose of a formulationcomprising bevacizumab to the patient once every two weeks wherein eachdose provides at least 5 mg/kg of bevacizumab, intravenously administera formulation comprising ABDNAZ at least once per week, and subject theglioma to radiation therapy at least twice per week. C For a duration ofat least 4 weeks, intravenously administer one dose of a formulationcomprising bevacizumab to the patient once every two weeks wherein eachdose provides at least 10 mg/kg of bevacizumab, intravenously administera formulation comprising ABDNAZ at least once per week, and subject theglioma to radiation therapy at least twice per week. D For a duration ofat least 6 weeks, intravenously administer one dose of a formulationcomprising bevacizumab to the patient once every three weeks whereineach dose provides at least 15 mg/kg of bevacizumab, intravenouslyadminister a formulation comprising ABDNAZ at least once per week, andsubject the glioma to radiation therapy at least twice per week. E For aduration of at least 4 weeks, intravenously administer one dose of aformulation comprising bevacizumab to the patient once every two weekswherein each dose provides at least 5 mg/kg of bevacizumab,intravenously administer a formulation comprising ABDNAZ at least twiceper week, and subject the glioma to radiation therapy at least twice perweek. F For a duration of at least 4 weeks, intravenously administer onedose of a formulation comprising bevacizumab to the patient once everytwo weeks wherein each dose provides at least 10 mg/kg of bevacizumab,intravenously administer a formulation comprising ABDNAZ at least twiceper week, and subject the glioma to radiation therapy at least twice perweek. G For a duration of at least 4 weeks, intravenously administer onedose of a formulation comprising bevacizumab to the patient once everytwo weeks wherein each dose provides at least 5 mg/kg of bevacizumab,intravenously administer a formulation comprising ABDNAZ at least twiceper week wherein each dose of the formulation comprising ABDNAZ providesfrom about 2.5 mg/m² to about 16.5 mg/m² of ABDNAZ, and subject theglioma to radiation therapy at least twice per week wherein each dose ofradiation therapy provides from about 1 Gy to about 3 Gy of radiation. HFor a duration of at least 4 weeks, intravenously administer one dose ofa formulation comprising bevacizumab to the patient once every two weekswherein each dose provides at least 10 mg/kg of bevacizumab,intravenously administer a formulation comprising ABDNAZ at least twiceper week wherein each dose of the formulation comprising ABDNAZ providesfrom about 2.5 mg/m² to about 16.5 mg/m² of ABDNAZ, and subject theglioma to radiation therapy at least twice per week wherein each dose ofradiation therapy provides from about 1 Gy to about 3 Gy of radiation. IFor a duration of at least 6 weeks, intravenously administer one dose ofa formulation comprising bevacizumab to the patient once every threeweeks wherein each dose provides at least 15 mg/kg of bevacizumab,intravenously administer a formulation comprising ABDNAZ at least twiceper week wherein each dose of the formulation comprising ABDNAZ providesfrom about 2.5 mg/m² to about 16.5 mg/m² of ABDNAZ, and subject theglioma to radiation therapy at least twice per week wherein each dose ofradiation therapy provides from about 1 Gy to about 3 Gy of radiation.

Further Characterization of the First, Second, and Third Methods

The methods may be further characterized according to, for example, thetype of glioma, patients for treatment, effects of the treatment,formulation of ABDNAZ, and form of ABDNAZ.

Type of Glioma

The methods may be further characterized according to the nature of theglioma. In certain embodiments, the glioma is an astrocytoma. In certainembodiments, the glioma is a malignant astrocytoma. In certainembodiments, the glioma is a glioblastoma. In certain embodiments, theglioma is a primary glioblastoma. In certain embodiments, the glioma isa secondary glioblastoma. In certain embodiments, the glioma is agliosarcoma. In certain embodiments, the glioma is an anaplastic gliomawith 1p/19q chromosomes intact. In certain embodiments, the glioma is ananaplastic oligodendroglioma or anaplastic oligoastrocytoma.

Patients for Treatment

The therapeutic method may be further characterized according to thepatient to be treated. In certain embodiments, the patient is an adulthuman. In certain other embodiments, the patient is a pediatric human.In certain embodiments, the patient has previously undergone surgery toremove at least some glioma tissue. In certain embodiments, the patienthas previously undergone surgery, to remove at least some glioma tissue,within six weeks prior to receiving the first dose of temozolomide underthe methods described herein using ABDNAZ as part of a combinationtherapy treatment.

Characterization of Treatment Effects

The therapeutic method may be further characterized according to theeffect of the treatment, such as (i) a reduction in the size of at leastone glioma tumors in the patient, and/or (ii) reduction in the number ofglioma tumor in the patient.

Accordingly, in certain embodiments, the therapeutic method ischaracterized by at least a 20% reduction in the size of at least oneglioma tumor in the patient. In certain other embodiments, there is atleast a 35% reduction in the size of at least one glioma tumor in thepatient. In certain other embodiments, there is at least a 50% reductionin the size of at least one glioma tumor in the patient.

In certain embodiments, there is at least a 20% reduction in the numberof glioma tumors in the patient. In certain other embodiments, there isat least a 35% reduction in the number of glioma tumors in the patient.In yet other embodiments, there is at least a 50% reduction in thenumber of glioma tumors in the patient.

Exemplary Formulations Comprising ABDNAZ

The method may be further characterized according to the formulationcomprising ABDNAZ that is administered to the patient. In certainembodiments, the formulation comprises ABDNAZ, dimethylacetamide, water,and a polyethylene glycol. In certain embodiments, the polyethyleneglycol has a number-average molecular weight of about 400 g/mol.

Form of ABDNAZ

In certain embodiments, the patient may be administered apharmaceutically acceptable salt of ABDNAZ.

Methods for Increasing the Amount of Temozolomide, Irinotecan, orBevacizumab in a Glioma

Another aspect of the invention provides a method for increasing theamount of temozolomide, irinotecan, or bevacizumab in a glioma. Themethod generally involves administering to a patient having a glioma (i)a formulation comprising temozolomide, irinotecan, or bevacizumab and(ii) an effective amount of a formulation comprising ABDNAZ so that theABDNAZ exerts physiological activity during a time period in which thetemozolomide, irinotecan, or bevacizumab is present in the patient, inorder to increase the amount of the temozolomide, irinotecan, orbevacizumab in the glioma. The method may be characterized according tothe percent increase in the amount of temozolomide, irinotecan, orbevacizumab in the glioma relative to the amount of temozolomide,irinotecan, or bevacizumab in the glioma following an identicaladministration procedure but absent administering said effective amountof a formulation comprising ABDNAZ.

Accordingly, one aspect of the invention provides a method of increasingthe amount of temozolomide in a glioma, where the method comprisesadministering to a patient having a glioma (i) a formulation comprisingtemozolomide and (ii) an effective amount of a formulation comprisingABDNAZ so that the ABDNAZ exerts physiological activity during a timeperiod in which the temozolomide is present in the patient, in order toincrease the amount of the temozolomide in the glioma.

Another aspect of the invention provides a method of increasing theamount of irinotecan in a glioma, where the method comprisesadministering to a patient having a glioma (i) a formulation comprisingirinotecan and (ii) an effective amount of a formulation comprisingABDNAZ so that the ABDNAZ exerts physiological activity during a timeperiod in which the irinotecan is present in the patient, in order toincrease the amount of the irinotecan in the glioma.

Another aspect of the invention provides a method of increasing theamount of bevacizumab in a glioma, where the method comprisesadministering to a patient having a glioma (i) a formulation comprisingbevacizumab and (ii) an effective amount of a formulation comprisingABDNAZ so that the ABDNAZ exerts physiological activity during a timeperiod in which the bevacizumab is present in the patient, in order toincrease the amount of the bevacizumab in the glioma.

Each of the foregoing methods may be further characterized according tofeatures described above in connection with the First, Second, and ThirdMethods. Additionally, the foregoing methods may be furthercharacterized according to the percent increase (e.g., at least a 5%,10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200% or300% increase) in the amount of temozolomide, irinotecan, or bevacizumabin the glioma relative to the amount of temozolomide, irinotecan, orbevacizumab in the glioma following an identical administrationprocedure but absent administering said effective amount of aformulation comprising ABDNAZ.

In certain embodiments, the formulation comprising ABDNAZ isadministered within about 4, 6, 8, 10, 12, 14, 16, 18, 20 or 24 hours ofadministration of a formulation comprising one of temozolomide,irinotecan, or bevacizumab.

Additional Method—Using Temozolomide

Another aspect of the invention provides a method of treating a gliomain a patient. The method comprises the steps of: (a) administering tothe patient in need thereof a therapeutically effective amount of aformulation comprising ABDNAZ, and (b) thereafter administering to thepatient in need thereof a therapeutically effective amount of aformulation comprising temozolomide; to treat the glioma. The method maybe further characterized by additional features, such as the dosingschedule and amount of temozolomide, dosing schedule of ABDNAZ, dosingamount for ABDNAZ, and/or any rest period/maintenance period, asdescribed above in connection with the First Method.

IV. Pharmaceutical Compositions

The invention provides pharmaceutical compositions comprising an activetherapeutic agent and one or more pharmaceutically acceptable carriers(additives) and/or diluents. In certain embodiments, the activetherapeutic agent is ABDNAZ, such that the invention provides apharmaceutical composition comprising ABDNAZ formulated together withone or more pharmaceutically acceptable carriers (additives) and/ordiluents. The a pharmaceutical composition may comprise ABDNAZ in atherapeutically effective amount. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in liquid form, including those adaptedfor the following: (1) oral administration, for example, drenches(aqueous or non-aqueous solutions or suspensions); and (2) parenteraladministration by, for example, subcutaneous, intramuscular, intravenousor epidural injection as, for example, a sterile solution or suspension,or sustained-release formulation.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect in at least a sub-population of cells in an animal ata reasonable benefit/risk ratio applicable to any medical treatment.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The amount of active ingredient which can be combined with a carriermaterial to produce a single dosage form will generally be that amountof the compound which produces a therapeutic effect. Generally, out ofone hundred percent, this amount will range from about 0.1 percent toabout ninety-nine percent of active ingredient, preferably from about 5percent to about 70 percent, most preferably from about 10 percent toabout 30 percent.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsugars, alcohols, antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms upon the subject compounds may be ensuredby the inclusion of various antibacterial and antifungal agents, forexample, paraben, chlorobutanol, phenol sorbic acid, and the like. Itmay also be desirable to include isotonic agents, such as sugars, sodiumchloride, and the like into the compositions. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption such as aluminummonostearate and gelatin.

The preparations of the present invention may be given, for example,orally or parenterally. The phrases “parenteral administration” and“administered parenterally” as used herein means modes of administrationother than enteral and topical administration, usually by injection, andincludes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intracapsular, intraorbital, intracardiac, intradermal,intraperitoneal, transtracheal, subcutaneous, subcuticular,intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternalinjection and infusion.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient which is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion ormetabolism of the particular compound being employed, the rate andextent of absorption, the duration of the treatment, other drugs,compounds and/or materials used in combination with the particularcompound employed, the age, sex, weight, condition, general health andprior medical history of the patient being treated, and like factorswell known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound which is the lowest dose effective toproduce a therapeutic effect. Such an effective dose will generallydepend upon the factors described above. Preferably, the compounds areadministered at about 0.01 mg/kg to about 200 mg/kg, more preferably atabout 0.1 mg/kg to about 100 mg/kg, even more preferably at about 0.5mg/kg to about 50 mg/kg. When the compounds described herein areco-administered with another agent (e.g., as sensitizing agents), theeffective amount may be less than when the agent is used alone.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms. Preferred dosing is one administrationper day.

The description above describes multiple aspects and embodiments of theinvention. The patent application specifically contemplates allcombinations and permutations of the aspects and embodiments.

V. Kits for Use in Medical Applications

Another aspect of the invention provides a kit for treating a glioma.The kit comprises: (i) a formulation comprising ABDNAZ, and (ii)instructions for treating a glioma according to procedures describedherein, such as (a) administering to the patient in need thereof atherapeutically effective amount of a formulation comprisingtemozolomide, and within about 2 days thereafter subjecting the gliomato radiation therapy; and (b) administering to the patient at least onedose of a therapeutically effective amount of a formulation comprisingABDNAZ within about 7 days of administration of a first dose of thetemozolomide; to treat the glioma. In an alternative embodiment, the kitcomprises: (i) a formulation comprising ABDNAZ, and (ii) instructionsfor treating a glioma that comprise (a) administering to the patient inneed thereof a therapeutically effective amount of a formulationcomprising irinotecan, and within about 7 days subjecting the glioma toradiation therapy; and (b) administering to the patient at least onedose of a therapeutically effective amount of a formulation comprisingABDNAZ within about 7 days of administration of a first dose of theirinotecan; to treat the glioma.

VI. Definitions

To facilitate an understanding of the present invention, a number ofterms and phrases are defined below.

The terms “a” and “an” as used herein mean “one or more” and include theplural unless the context is inappropriate.

As used herein, the term “patient” refers to organisms to be treated bythe methods of the present invention. Such organisms are preferablymammals (e.g., murines, simians, equines, bovines, porcines, canines,felines, and the like), and more preferably humans.

As used herein, the term “effective amount” refers to the amount of acompound (e.g., a compound of the present invention) sufficient toeffect beneficial or desired results. An effective amount can beadministered in one or more administrations, applications or dosages andis not intended to be limited to a particular formulation oradministration route.

As used herein, the term “treating” includes any effect, e.g.,lessening, reducing, modulating, ameliorating or eliminating, thatresults in the improvement of the condition, disease, disorder, and thelike, or ameliorating a symptom thereof.

As used herein, the terms “alleviate” and “alleviating” refer toreducing the severity of the condition, such as reducing the severityby, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,or 95%.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent with a carrier, inert or active, makingthe composition especially suitable for diagnostic or therapeutic use invivo or ex vivo.

As used herein, the term “pharmaceutically acceptable carrier” refers toany of the standard pharmaceutical carriers, such as a phosphatebuffered saline solution, water, emulsions (e.g., such as an oil/wateror water/oil emulsions), and various types of wetting agents. Thecompositions also can include stabilizers and preservatives. Forexamples of carriers, stabilizers and adjuvants, see, for example,Martin, Remington's Pharmaceutical Sciences, 15th Ed., Mack Publ. Co.,Easton, Pa. [1975]).

As used herein, the term “pharmaceutically acceptable salt” refers toany pharmaceutically acceptable salt (e.g., acid or base) of a compoundof the present invention which, upon administration to a subject, iscapable of providing a compound of this invention or an activemetabolite or residue thereof. As is known to those of skill in the art,“salts” of the compounds of the present invention may be derived frominorganic or organic acids and bases. Examples of acids include, but arenot limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric,fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic,ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic,benzenesulfonic acid, and the like. Other acids, such as oxalic, whilenot in themselves pharmaceutically acceptable, may be employed in thepreparation of salts useful as intermediates in obtaining the compoundsof the invention and their pharmaceutically acceptable acid additionsalts.

Examples of bases include, but are not limited to, alkali metal (e.g.,sodium) hydroxides, alkaline earth metal (e.g., magnesium) hydroxides,ammonia, and compounds of formula NW₄ ⁺, wherein W is C₁₋₄ alkyl, andthe like.

Examples of salts include, but are not limited to: acetate, adipate,alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate,citrate, camphorate, camphorsulfonate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate,glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate,pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate,succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like.Other examples of salts include anions of the compounds of the presentinvention compounded with a suitable cation such as Na⁺, NH₄ ⁺, and NW₄⁺ (wherein W is a C₁₋₄ alkyl group), and the like.

For therapeutic use, salts of the compounds of the present invention arecontemplated as being pharmaceutically acceptable. However, salts ofacids and bases that are non-pharmaceutically acceptable may also finduse, for example, in the preparation or purification of apharmaceutically acceptable compound.

Throughout the description, where compositions are described as having,including, or comprising specific components, or where processes andmethods are described as having, including, or comprising specificsteps, it is contemplated that, additionally, there are compositions ofthe present invention that consist essentially of, or consist of, therecited components, and that there are processes and methods accordingto the present invention that consist essentially of, or consist of, therecited processing steps.

As a general matter, compositions specifying a percentage are by weightunless otherwise specified. Further, if a variable is not accompanied bya definition, then the previous definition of the variable controls.

EXAMPLES

The invention now being generally described, will be more readilyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention, and are not intended to limit the invention.

Example 1—Treatment of Human Patients Suffering from a Glioblastoma orMalignant Glioma

Adult human patients having a gliobastoma or malignant glioma are to betreated using a combination of ABDNAZ, temozolomide, and radiationtherapy (RT) according to the treatment protocol described below.

Treatment Protocol Upfront Therapy

Patients are to receive oral temozolomide 75 mg/m² daily for 6 weeks andundergo conformal or intensity-modulated radiotherapy (60 Gy in 2 Gyfractions) 5 days a week for 6 weeks. Patients are also to receiveABDNAZ by intravenous administration twice weekly for 6 weeksadministered at one of four dose levels (2.0 mg/m², 8 mg/m², 17 mg/m²,or 29 mg/m²) beginning concurrently with temozolomide and radiotherapy.Cohorts of 3-6 patients will receive escalating doses of ABDNAZ untilthe maximum feasible dose (MFD) or maximum tolerated dose (MTD) isdetermined. The MTD is defined as the dose preceding that at which 2 of3 or 2 of 6 subjects experience dose-limiting toxicity.

Maintenance Therapy

Beginning 28 days after completion of radiotherapy, patients are toreceive oral temozolomide [150 mg/m² orally daily for 5 days duringcycle 1 (28 days) and 200 mg/m² orally daily for 5 days every 28 daysfor cycles 2-6] and once weekly ABDNAZ at 2 mg/m² on days 1-28.Treatment will repeat every 4 weeks for up to 12 courses in the absenceof disease progression or unacceptable toxicity.

Patient Evaluation Procedure

Patients are to be evaluated by a radiation oncologist and medicaloncologist prior to the start of any treatment. Efficacy of the therapymay be evaluated according to procedure described below.

Pre-Treatment Evaluation (within 14±7 Days Prior to First Dose of StudyDrug)

Patients may be evaluated using the following criteria prior totreatment with the therapy to assess nature and extent of thegliobastoma or malignant glioma:

-   -   GAD-enhanced DCE-MRI    -   Contrast-enhanced MRI and/or CT    -   Complete history and general physical exam    -   Detailed neurological examination    -   Safety labs: CBC with differential, comprehensive chemistry        panel, liver function tests    -   Karnofsky performance status    -   Documentation of primary tumor histology    -   Documentation of steroid and anticonvulsant doses and any other        concomitant medications. The newer antiepilieptic drugs        (levetiracetam, pregabalin, lamotrigine, lacosamide, topiramate)        are generally preferred over carbamazepine, phenytoin,        phenobarbital, or valproic acid, which stimulate the hepatic        cytochrome P450 (CYP) system.    -   FACT-BR, Barthel Index of ADLs, and neurocognitive function        (HVLT-R, COWA, and Trail Making A and B) will be evaluated

Evaluation During Radiation Therapy and ABDNAZ Therapy

Patients may be evaluated using the following criteria during treatmentwith the therapy to assess nature and extent of the gliobastoma ormalignant glioma:

-   -   Clinical visit once per week to include neurological        examination, history and physical examination and clinical        laboratory testing    -   GAD-enhanced DCE-MRI on days 9±1 day and 41±1 day    -   Collection of concomitant medications at each clinic visit    -   Collection of adverse events (AEs) at each clinic visit    -   Skin within the treatment portal shall be examined once per week        during radiation treatment        Evaluation after Completion of Radiation Therapy and ABDNAZ        Therapy

Patients may be evaluated using the following criteria after treatmentwith the therapy to assess nature and extent of the gliobastoma ormalignant glioma:

-   -   Patients are to be evaluated 2 weeks after completion of        radiation therapy and ABDNAZ treatment with neurological exam,        history and symptom-directed physical examination, and clinical        laboratory testing    -   During the maintenance phase, patients are to be evaluated on        Day 1±2 days of each 4-week cycle. Each evaluation will consist        of neurological examination, history and symptom-directed        physical examination, ECOG performance status, FACT-BR, Barthel        Index of ADLs, and neurocognitive function    -   Safety labs every 4 weeks on Day 1±2 days for Cycles 2+(CBC with        differential, comprehensive chemistry panel, liver function        tests). Radiologic evaluation will occur every 8 weeks as per        Standard of Care    -   Patients are to be evaluated with contrast-enhanced MRI w/GAD        prior to starting Cycle 1 and at least every 2 months (after        every 2 cycles) of ABDNAZ/temozolomide as well as neurocognitive        testing at 4 months±7 days (1 month=4 weeks or 28 days) after        completion of radiation therapy/ABDNAZ    -   Collection of concomitant medications at each clinic visit    -   Collection of adverse events (AEs) at each clinic visit

General Features

During Upfront Therapy and Maintenance Therapy described above,temozolomide is to be administered continuously from day 1 ofradiotherapy to the last day of radiation at a daily oral dose of 75mg/m² for a maximum of 49 days (to allow for treatments interruptions,for example, over holidays). The drug will be administered at night justbefore bedtime. The dose will be determined using actual body surfacearea (BSA) as defined by the method of DuBois and DuBois. Capsules oftemozolomide are available in 5, 20, 100, 140, 180, and 250 mg. Thedaily dose is to be rounded to the nearest 5 mg. Patients are to beinstructed to swallow the capsules whole, in rapid succession, withoutchewing them. If vomiting occurs during the course of treatment, nore-dosing of the patient is allowed before the next scheduled dose. Thecapsules should be taken on an empty stomach, therefore a minimum of 2hours after eating and with no food consumption for at least 1 hourafter temozolomide administration. Water is allowed during the fastperiod. Administration of the higher dosing regimen during themaintenance phase of the protocol should also be at night. However,prophylaxis with a 5-HT3 antagonist is recommended prior toadministration of the first few temozolomide doses and may beadministered orally 30 to 60 minutes before temozolomide treatment.

For radiation therapy, 2 Gy of radiation is to be given daily 5 days perweek for a total of 60 Gy over 6 weeks. All fields shall be treatedduring each treatment session. Doses are specified such that at least95% of the planning treatment volume (PTV) shall receive 100% of theprescribed dose; Dose Volume Histograms (DVHs) may be necessary to makethis selection. Radiation therapy will be delivered using standard doseand fractionation as described below:

-   -   Simulation, Immobilization: Patients are to be treated in the        supine position. Adequate immobilization and reproducibility of        position will be ensured using thermoplastic mask.    -   Target: The target volume for both the initial volume and the        conedown volume shall be based on CT/MRI. This initial target        volume shall include the contrast-enhancing lesion and        surrounding edema (if it exists) demonstrated on CT/MRI plus a        margin. If no surrounding edema is present, the initial target        volume should include the contrast enhancing lesion plus a        margin. The initial target volume will be treated to 46 Gy in 23        fractions. After 46 Gy, the tumor volume for the conedown        treatment should include the contrast enhancing lesion (without        edema) on the CT/MRI scan plus a margin. Isodose distributions        for the initial target volume and the conedown target volume are        required on all patients, including those treated with parallel        opposed fields. The inhomogeneity within the target volume shall        be kept to 10%. The minimum dose to the target volume should be        kept within 10% of the dose at the center of the volume.    -   Dose Limitations: The lens and cervical spine should be shielded        from the direct beam at all times. When possible to do without        shielding gross tumor, attempts should be made to limit the dose        to the optic chiasm to 54 Gy, the retina of at least one eye        (but preferably both) to 50 Gy, and the brainstem to 60 Gy. When        the optic chiasm must be included in the full dose, then there        may be a finite unknown risk of developing blindness.    -   Dose Specification: Radiotherapy is to be delivered with a daily        fraction size of 2.0 Gy per fraction given 5 days a week for a        total of 60 Gy over 6 weeks.    -   Technique: Treatment will be delivered with megavoltage machines        with a minimum energy of 6 MV photons. All fields will be        treated during each treatment session. Treatment plans may        include opposed lateral fields, a wedge pair of fields,        rotation, or multiple field techniques. Intensity-modulated        inverse-planned approaches are permitted. Any of the methods of        IMRT (including tomotherapy) may be used, subject to protocol        localization and dosimetry constraints. CT-based treatment        planning is desirable to assure accuracy in the selection of        field arrangements. MRI-fusion for accurate target delineation        is recommended.

Example 2—Treatment of Human Patients Suffering from a Glioblastoma orMalignant Glioma

Adult human patients having a gliobastoma or malignant glioma are to betreated using a combination of ABDNAZ (by intravenous administration ata dose ranging from 0.5 mg to 4.0 mg), temozolomide, and radiationtherapy (RT) according to the treatment protocol described below.

Treatment Protocol Upfront Therapy

Patients are to receive oral temozolomide 75 mg/m² daily for 6 weeks andundergo conformal or intensity-modulated radiotherapy (60 Gy in 2 Gyfractions) 5 days a week for 6 weeks. Patients are also to receiveABDNAZ by intravenous administration once weekly for 6 weeksadministered at one of four dose levels (0.5 mg, 1.0 mg, 2.0 mg, or 4.0mg) beginning concurrently with temozolomide and radiotherapy. Cohortsof 3-6 patients will receive escalating doses of ABDNAZ until themaximum feasible dose (MFD) or maximum tolerated dose (MTD) isdetermined. The MTD is defined as the dose preceding that at which 2 of3 or 2 of 6 subjects experience dose-limiting toxicity. Administrationof temozolomide is no sooner than 4 hours after administration ofABDNAZ. ABDNAZ is preferably administered within three hours prior toadministration of radiation therapy.

Maintenance Therapy

Beginning 4-6 weeks after completion of radiotherapy, patients are toreceive oral temozolomide [150 mg/m² orally daily for on days 1-5 duringcycle 1 (28 days) and 200 mg/m² orally daily on days 1-5 during cycles2-6 (where each cycle is 28 days)] and once weekly ABDNAZ by intravenousadministration at a dose ranging from 0.5 mg to 4.0 mg for six months.Administration of temozolomide is no sooner than 4 hours afteradministration of ABDNAZ.

Patient Evaluation Procedure

Patients are to be evaluated by a neuro-oncologist, radiationoncologist, or medical oncologist prior to the start of any treatment.Efficacy of the therapy may be evaluated according to proceduredescribed below.

Pre-Treatment Evaluation (within 14±7 Days Prior to First Dose of StudyDrug)

Patients may be evaluated using the following criteria prior totreatment with the therapy to assess nature and extent of thegliobastoma or malignant glioma:

-   -   Contrast-enhanced MRI    -   Complete history and general physical exam    -   Detailed neurological examination    -   Safety labs: CBC with differential, comprehensive chemistry        panel with liver function tests    -   Karnofsky performance status    -   Documentation of primary tumor histology    -   Documentation of steroid and anticonvulsant doses and any other        concomitant medications. The newer antiepilieptic drugs        (levetiracetam, pregabalin, lamotrigine, lacosamide, topiramate)        are generally preferred over carbamazepine, phenytoin,        phenobarbital, or valproic acid, which stimulate the hepatic        cytochrome P450 (CYP) system.

Evaluation During Radiation Therapy and ABDNAZ Therapy

Patients may be evaluated using the following criteria during treatmentwith the therapy to assess nature and extent of the gliobastoma ormalignant glioma:

-   -   Clinical visit once per week to include neurological        examination, history and physical examination and clinical        laboratory testing    -   Documentation of concomitant medications at each clinic visit    -   Collection of adverse events (AEs) at each clinic visit    -   Skin within the treatment portal shall be examined once per week        during radiation treatment    -   Serum magnesium test during week 4        Evaluation after Completion of Radiation Therapy and ABDNAZ        Therapy

Patients may be evaluated using the following criteria after treatmentwith the therapy to assess nature and extent of the gliobastoma ormalignant glioma:

-   -   Patients are to be evaluated 3-5 weeks after completion of        radiation therapy and ABDNAZ treatment with neurological exam,        history and symptom-directed physical examination, adverse        events, concomitant medications, and clinical laboratory testing    -   During the maintenance phase, patients are to be evaluated on        Day 1±2 days of each 4-week cycle. Each evaluation will consist        of neurological examination, history and symptom-directed        physical examination, and KPS/ECOG performance status    -   Safety labs every 4 weeks on Day 1±2 days for Cycles 2+(CBC with        differential, comprehensive chemistry panel, liver function        tests). Radiologic evaluation will occur every 8 weeks as per        Standard of Care    -   Patients are to be evaluated with contrast-enhanced MRI w/GAD        prior to starting Cycle 1 and at least every 2 months (after        every 2 cycles) of ABDNAZ/temozolomide as well as neurocognitive        testing at 4 months±7 days (1 month=4 weeks or 28 days) after        completion of radiation therapy/ABDNAZ    -   Collection of concomitant medications at each clinic visit    -   Collection of adverse events (AEs) at each clinic visit

General Features

During Upfront Therapy and Maintenance Therapy described above,temozolomide is to be administered continuously from day 1 ofradiotherapy to the last day of radiation at a daily oral dose of 75mg/m² for a maximum of 49 days (to allow for treatments interruptions,for example, over holidays). The drug will be administered at night justbefore bedtime. The dose will be determined using actual body surfacearea (BSA) as defined by the method of DuBois and DuBois. Capsules oftemozolomide are available in 5, 20, 100, 140, 180, and 250 mg. Thedaily dose is to be rounded to the nearest 5 mg. Patients are to beinstructed to swallow the capsules whole, in rapid succession, withoutchewing them. If vomiting occurs during the course of treatment, nore-dosing of the patient is allowed before the next scheduled dose. Thecapsules should be taken on an empty stomach, therefore a minimum of 2hours after eating and with no food consumption for at least 1 hourafter temozolomide administration. Water is allowed during the fastperiod. Administration of the higher dosing regimen during themaintenance phase of the protocol should also be at night. However,prophylaxis with a 5-HT3 antagonist is recommended prior toadministration of the first few temozolomide doses and may beadministered orally 30 to 60 minutes before temozolomide treatment.

For radiation therapy, 2 Gy of radiation is to be given daily 5 days perweek for a total of 60 Gy over 6 weeks. All fields shall be treatedduring each treatment session. Doses are specified such that at least95% of the planning treatment volume (PTV) shall receive 100% of theprescribed dose; Dose Volume Histograms (DVHs) may be necessary to makethis selection. Radiation therapy will be delivered using standard doseand fractionation as described below:

-   -   Simulation, Immobilization: Patients are to be treated in the        supine position. Adequate immobilization and reproducibility of        position will be ensured using thermoplastic mask.    -   Target: The target volume for both the initial volume and the        conedown volume shall be based on CT/MRI. This initial target        volume shall include the contrast-enhancing lesion and        surrounding edema (if it exists) demonstrated on CT/MRI plus a        margin. If no surrounding edema is present, the initial target        volume should include the contrast enhancing lesion plus a        margin. The initial target volume will be treated to 46 Gy in 23        fractions. After 46 Gy, the tumor volume for the conedown        treatment should include the contrast enhancing lesion (without        edema) on the CT/MRI scan plus a margin. Isodose distributions        for the initial target volume and the conedown target volume are        required on all patients, including those treated with parallel        opposed fields. The inhomogeneity within the target volume shall        be kept to 10%. The minimum dose to the target volume should be        kept within 10% of the dose at the center of the volume.    -   Dose Limitations: The lens and cervical spine should be shielded        from the direct beam at all times. When possible to do without        shielding gross tumor, attempts should be made to limit the dose        to the optic chiasm to 54 Gy, the retina of at least one eye        (but preferably both) to 50 Gy, and the brainstem to 60 Gy. When        the optic chiasm must be included in the full dose, then there        may be a finite unknown risk of developing blindness.    -   Dose Specification: Radiotherapy is to be delivered with a daily        fraction size of 2.0 Gy per fraction given 5 days a week for a        total of 60 Gy over 6 weeks.    -   Technique: Treatment will be delivered with megavoltage machines        with a minimum energy of 6 MV photons. All fields will be        treated during each treatment session. Treatment plans may        include opposed lateral fields, a wedge pair of fields,        rotation, or multiple field techniques. Intensity-modulated        inverse-planned approaches are permitted. Any of the methods of        IMRT (including tomotherapy) may be used, subject to protocol        localization and dosimetry constraints. CT-based treatment        planning is desirable to assure accuracy in the selection of        field arrangements. MRI-fusion for accurate target delineation        is recommended.

Example 3—Administration of ABDNAZ and Temozolomide to a GBM14 TMZ-SPatient-Derived Cell Line and a GBM14 TMZ-R Patient-Derived Cell Line

Temozolomide-sensitive human GBM14 (GBM14 TMZ-S) patient derived cellline cells were treated with one of the following (i) ABDNAZ, (ii)temozolomide, or (iii) ABDNAZ and temozolomide. In a separateexperiment, temozolomide-resistant human GBM14 (GBM14 TMZ-R) patientderived cell line cells were treated with one of the following (i)ABDNAZ, (ii) temozolomide, or (iii) ABDNAZ and temozolomide.Experimental procedures and results are provided below.

Part I—Procedures

In a first experiment, temozolomide-sensitive human GBM14 (GBM14 TMZ-S)patient derived cell line cells were treated with one of the following(i) ABDNAZ at a concentration of 2 μM, (ii) temozolomide at aconcentration of 100 μM, or (iii) ABDNAZ (at a concentration of 2 μM)and temozolomide (at a concentration of 100 μM). WST-1 assay was used toassess the effect of treatment on cell viability over the time of threedays. The presence of live cells is detected by absorbance at 440 nm inthe assay. Data are presented as the mean±3 standard deviations fromthree separate performances of the experiment.

In a second experiment, temozolomide-resistant human GBM14 (GBM14 TMZ-R)patient derived cell line cells were treated with one of the following(i) ABDNAZ at a concentration of 2 μM, (ii) temozolomide at aconcentration of 100 μM, or (iii) ABDNAZ (at a concentration of 2 μM)and temozolomide (at a concentration of 100 μM). The presence of livecells is detected by absorbance at 440 nm in the assay. WST-1 assay wasused to assess the effect of treatment on cell viability over the timeof three days. Data are presented as the mean±3 standard deviations fromthree separate performances of the experiment.

Part II—Results

Results are presented in FIG. 1 showing response from the GBM14 TMZ-Spatient derived cell line and FIG. 2 showing response from the GBM14TMZ-R patient derived cell line. As indicated by data in FIGS. 1 and 2,administration of ABDNAZ and temozolomide together produced a greateramount of cell death as measured by lack of absorbance at 440 nm.

Example 4—Administration of ABDNAZ and Temozolomide to NSCG Cells InVitro

Cells from temozolomide-resistant murine EGFRvIII/Arfko neural stemcell-derived glioma (NSCG) cell line were treated with one of thefollowing: (i) ABDNAZ (RRx-001), (ii) temozolomide (TMZ), or (iii)ABDNAZ and temozolomide. Experimental procedures and results areprovided below.

Part I—Procedures

Cells from temozolomide-resistant murine EGFRvIII/Arfko neural stemcell-derived glioma (NSCG) cell line were treated with one of thefollowing (i) ABDNAZ at a concentration of 2 μM, (ii) temozolomide at aconcentration of 100 μM, or (iii) ABDNAZ (at a concentration of 2 μM)and temozolomide (at a concentration of 100 μM). WST-1 assay was used toassess the effect of treatment on cell viability over the time of fivedays. The presence of live cells is detected by absorbance at 440 nm inthe assay. Data are presented as the mean±3 standard deviations fromthree separate performances of the assay.

Part II—Results

Results are presented in FIG. 3, where the data indicate thatadministration of ABDNAZ and temozolomide together produced a greateramount of cell death as measured by lack of absorbance at 440 nm. Dataare presented as the mean±3 standard deviations from three separateperformances of the assay.

Example 5—Administration of ABDNAZ and Temozolomide to Mice Implantedwith NSCG Tumors

Mice implanted with temozolomide-resistant EGFRvIII/Arfko GBM (NSCG)cells were treated with (i) ABDNAZ (RRx-001, 10 mg/kg, intraperitonealinjection), (ii) temozolomide (TMZ)(40 mg/kg, oral gavage), or (iii)ABDNAZ (10 mg/kg, intraperitoneal injection) and temozolomide (40 mg/kg,oral gavage). Experimental procedures and results are provided below.

Part I—Procedures

NSCG tumors were intracranially injected into FVBN mice. Mice to betreated were treated with one of the following (i) ABDNAZ (10 mg/kg,intraperitoneal injection), (ii) temozolomide (40 mg/kg, oral gavage),or (iii) ABDNAZ (10 mg/kg, intraperitoneal injection) and temozolomide(40 mg/kg, oral gavage). In a first experiment, treatment (i.e.,administration of ABDNAZ (RRx-001), temozolomide (TMZ), or both ABDNAZand temozolomide) was started five days after intracranial injection ofNSCG cells. The control population of mice contained ten mice, the groupreceiving only ABDNAZ had twelve mice, the group receiving onlytemozolomide had twelve mice, and the group receiving both ABDNAZ andtemozolomide had sixteen mice. Mouse survival was monitored over time.

In a second experiment, treatment (i.e., administration of ABDNAZ(RRx-001), temozolomide (TMZ), or both ABDNAZ and temozolomide) wasstarted ten days after intracranial injection of NSCG cells. The controlpopulation of mice contained three mice, the group receiving only ABDNAZhad five mice, the group receiving only temozolomide had four mice, andthe group receiving both ABDNAZ and temozolomide had six mice. Mousesurvival was monitored over time.

Part II—Results

Results are presented in FIGS. 4 and 5 and Tables 1 and 2 below. FIG. 4is a graph showing percent survival of subjects over time, in theexperiment in which treatment (i.e., administration of ABDNAZ (RRx-001),temozolomide (TMZ), or both ABDNAZ and temozolomide) was started fivedays after intracranial injection of NSCG cells. The abbreviation “CTL”in FIG. 4 refers to control subjects. Table 1 shows median survival time(in days) observed in the experiment in which treatment (i.e.,administration of ABDNAZ (RRx-001), temozolomide (TMZ), or both ABDNAZand temozolomide) was started five days after intracranial injection ofNSCG cells.

FIG. 5 is a graph showing percent survival of subjects over time, in theexperiment in which treatment (i.e., administration of ABDNAZ (RRx-001),temozolomide (TMZ), or both ABDNAZ and temozolomide) was started tendays after intracranial injection of NSCG cells. The abbreviation “CTL”in FIG. 5 refers to control subjects. Table 2 shows median survival time(in days) observed in the experiment in which treatment (i.e.,administration of ABDNAZ (RRx-001), temozolomide (TMZ), or both ABDNAZand temozolomide) was started ten days after intracranial injection ofNSCG cells.

TABLE 1 Subjects Population Population Population Receiving ReceivingOnly Receiving Both Control Only TMZ ABDNAZ ABDNAZ & TMZ Median 22.222.3 25.3 30.4 Survival Time (days) (ns) (p = 0.072) (p = 0.003)

TABLE 2 Subjects Population Population Population Receiving ReceivingOnly Receiving Both Control Only TMZ ABDNAZ ABDNAZ & TMZ Median 22.122.3 22.5 27.2 Survival Time (days) (ns) (ns) (p = 0.174)

Example 6—Administration of ABDNAZ and Temozolomide to Mice Implantedwith NSCG Tumors

Mice implanted with TMZ-resistant EGFRvIII/Arfko GBM tumors (NSCGtumors) were treated with ABDNAZ (RRx-001), temozolomide (TMZ), or bothABDNAZ and temozolomide. Experimental procedures and results areprovided below.

Part I—Procedures

NSCG tumors were orthotopically implanted in FVBN mice. Then, the micewere treated with one of the following (i) ABDNAZ (10 mg/kg,intraperitoneal injection), (ii) temozolomide (40 mg/kg, oral gavage),or (iii) ABDNAZ (10 mg/kg, intraperitoneal injection) and temozolomide(40 mg/kg, oral gavage). Percent apoptosis of CC3+ cells was evaluated.

Part II—Results

The amount of CC3+ cell apoptosis detected in the assay is shown in FIG.6. As indicated by data in FIG. 6, administration of ABDNAZ andtemozolomide together produced the highest percentage of CC3+ cellapoptosis. The abbreviation “CTL” refers to control mice; the symbol ***refers to p<0.001.

Example 7—Administration of ABDNAZ and Temozolomide to Mice Implantedwith Brain Tumors

Mice implanted with brain tumors were treated with ABDNAZ, temozolomide(TMZ), or both ABDNAZ and temozolomide. Experimental procedures andresults are provided below.

Part I—Procedures

Brain tumors were orthotopically implanted in the brain of mice. Then,the mice were treated with one of the following (i) ABDNAZ, (ii)temozolomide, or (iii) ABDNAZ followed by administration of temozolomide4 hours later. ABDNAZ (10 mg/kg) was administered via tail vein 4 hoursbefore any temozolomide (20 mg/kg) via tail vein once weekly for 4weeks. Twenty-four hours after temozolomide administration,tumor-bearing mice were euthanized and brain tumor tissue was collectedand dissected. Temozolomide levels in tumor tissues were measured.

Part II—Results

The concentration of temozolomide (TMZ) detected in brain tumor tissueof the mice is shown in FIG. 7. As indicated by data in FIG. 7,administration of ABDNAZ prior to administration of temozolomideresulted in a greater concentration of temozolomide in the brain tumortissue.

Example 8—Administration of ABDNAZ and Irinotecan to Mice Implanted withHuman Glioblastoma GBM43 Brain Tumors

Mice implanted with human glioblastoma GBM43 brain tumors were treatedwith ABDNAZ, irinotecan, or both ABDNAZ and irinotecan. Experimentalprocedures and results are provided below.

Part I—Procedures

Human glioblastoma GBM43 tumors were orthotopically implanted in thebrain of immunocompromised mice. Then, the mice were treated with one ofthe following (i) ABDNAZ, (ii) irinotecan, or (iii) ABDNAZ followed byadministration of irinotecan 4 hours later. Twenty-four hours afteririnotecan administration, tumor-bearing mice were euthanized and braintumor tissue was collected and dissected. Irinotecan levels in tumortissues were measured.

Part II—Results

The concentration of irinotecan detected in brain tumor tissue of themice is shown in FIG. 8. As indicated by data in FIG. 8, administrationof ABDNAZ prior to administration of irinotecan resulted in a greaterconcentration of irinotecan in the brain tumor tissue.

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents and scientificarticles referred to herein is incorporated by reference for allpurposes.

EQUIVALENTS

The invention may be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof. The foregoingembodiments are therefore to be considered in all respects illustrativerather than limiting the invention described herein. Scope of theinvention is thus indicated by the appended claims rather than by theforegoing description, and all changes that come within the meaning andrange of equivalency of the claims are intended to be embraced therein.

1. A method of treating a glioma in a patient, comprising the steps of:(a) administering to the patient in need thereof a therapeuticallyeffective amount of a formulation comprising temozolomide, and withinabout 2 days thereafter subjecting the glioma to radiation therapy; and(b) administering to the patient at least one dose of a therapeuticallyeffective amount of a formulation comprising ABDNAZ within about 7 daysof administration of a first dose of the temozolomide; to treat theglioma; wherein each dose of the formulation comprising ABDNAZ isadministered to the patient by intravenous infusion providing ABDNAZ inan amount of from about 0.1 mg to about 20 mg.
 2. The method of claim 1,wherein the formulation comprising temozolomide is administered dailyfor at least 2 weeks. 3.-5. (canceled)
 6. The method of claim 1, whereinthe patient receives temozolomide by oral administration at a daily doseof at least 50 mg/m². 7.-8. (canceled)
 9. The method of claim 1, whereinfor a duration of at least 2 weeks following administration of the firstdose of temozolomide, the patient receives at least one dose each weekof a therapeutically effective amount of a formulation comprisingABDNAZ. 10.-11. (canceled)
 12. The method of claim 1, wherein two dosesof a therapeutically effective amount of a formulation comprising ABDNAZare administered within about 7 days of administration of the first doseof temozolomide.
 13. The method of claim 1, wherein for a duration of atleast 2 weeks following administration of the first dose oftemozolomide, the patient receives two doses each week of atherapeutically effective amount of a formulation comprising ABDNAZ.14.-35. (canceled)
 36. The method of claim 1, wherein the formulationcomprising ABDNAZ contains dimethylacetamide, water, and a polyethyleneglycol.
 37. The method of claim 36, wherein the polyethylene glycol hasa number-average molecular weight of about 400 g/mol.
 38. The method ofclaim 1, wherein the glioma is subjected to radiation therapy once perday for at least 3 days within a 7 day period following administrationof the first dose of temozolomide.
 39. The method of claim 1, whereinthe glioma is subjected to radiation therapy once per day for at least 5days within a 7 day period following administration of the first dose oftemozolomide. 40.-47. (canceled)
 48. The method of claim 1, wherein whenradiation therapy is administered to the glioma, the amount of radiationprovided to the glioma on the day of administering the radiation therapyis from about 1 Gy to about 3 Gy. 49.-52. (canceled)
 53. The method ofclaim 1, wherein after receiving temozolomide for a duration of 6 weeks,the patient does not receive any of temozolomide, radiation therapy, orABDNAZ for a duration of at least about 2 weeks. 54.-56. (canceled) 57.The method of claim 1, wherein after receiving temozolomide for aduration of 6 weeks, the patient does not receive any of temozolomide,radiation therapy, or ABDNAZ for a duration of at least about 4 weeks.58.-68. (canceled)
 69. A method of treating a glioma in a patient,comprising the steps of: (a) administering to the patient in needthereof a therapeutically effective amount of a formulation comprisingirinotecan, and within about 7 days subjecting the glioma to radiationtherapy; and (b) administering to the patient at least one dose of atherapeutically effective amount of a formulation comprising ABDNAZwithin about 7 days of administration of a first dose of the irinotecan;to treat the glioma; wherein each dose of the formulation comprisingABDNAZ is administered to the patient by intravenous infusion providingABDNAZ in an amount of from about 0.1 mg to about 20 mg. 70.-107.(canceled)
 108. The method of claim 69, wherein the glioma is anastrocytoma.
 109. The method of claim 69, wherein the glioma is amalignant astrocytoma.
 110. The method of claim 69, wherein the gliomais a glioblastoma. 111.-112. (canceled)
 113. The method of claim 69,wherein the patient is an adult human. 114.-115. (canceled)
 116. Themethod of claim 69, wherein there is at least a 35% reduction in thesize of at least one glioma tumor in the patient. 117.-118. (canceled)119. The method of claim 69, wherein there is at least a 35% reductionin the number of glioma tumors in the patient.
 120. (canceled)